METHODS FOR GENOTYPING MATURE COTTON FIBERS AND TEXTILES

• US 20100279282A1
• Filed: 11/12/2008
• Published: 11/04/2010
• Est. Priority Date: 11/12/2008
• Status: Active Grant

• Current Assignee: APDN (B.V.I.) Inc. (Applied DNA Sciences Incorporated)
• Original Assignee: APDN (B.V.I.) Inc. (Applied DNA Sciences Incorporated)
• Inventors: Suh Kin So, Stephane, Liang, Ming-Hwa
• Granted Patent: US 8,940,485 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/13 Plant traits
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

Methods for distinguishing between cotton cultivars of a specific species by analyzing a sample of mature cotton fibers from raw cotton materials or from textile goods are disclosed. DNA is extracted from the mature cotton fiber sample and subjected to PCR techniques which enable the identification of the cultivar of a particular cotton species utilized in the textile or cotton material of interest.

Claims

1. A method for identifying cotton cultivar comprising:

collecting cotton fibers from a sample;

extracting genomic DNA from the cotton fibers collected;

amplifying said DNA with PCR-based techniques; and

analyzing the amplified product to distinguish between a first cotton cultivar and a second cotton cultivar.

2. The method of claim 1 wherein first cotton cultivar is from the species G. barbadense.

3. The method of claim 2 wherein the second cotton cultivar is from the G. barbadense.

4. The method of claim 1 wherein the sample is a textile item selected from a group consisting of raw cotton, ginned cotton, yarn blended with cotton, and fabrics blended with cotton.

5. The method of claim 1 wherein the amplifying further comprises;

using at least one set of specific primers which targets sequence polymorphism between the first cotton cultivar and the second cotton cultivar.

6. The method of claim 3 wherein amplifying further comprises;

using at least one set of specific primers which targets sequence polymorphism between G. barbadense cultivars.

7. The method of claim 5 wherein the targeted sequence of the first cotton cultivar and the second cotton cultivar have different detectable physical characteristics.

8. The method of claim 7 wherein the detectable physical characteristic of the targeted sequence is a different sequence length for the first cotton cultivar compared the second cotton cultivar.

9. The method of claim 1 wherein the genomic DNA extracted from the cotton fibers comprises chloroplast DNA.

10. The method of claim 1 wherein the genomic DNA extracted from the cotton fibers comprises mitochondrial DNA.

11. The method of claim 1 wherein the genomic DNA extracted from the cotton fibers comprises nuclear DNA.

12. The method of claim 6 wherein the detectable physical characteristic of the targeted sequence is a different sequence composition.

13. The method of claim 6 wherein the detectable physical characteristic of the targeted sequence is a different sequence length.

14. The method of claim 13 wherein the analyzing of the amplified products comprises detecting the size of the amplified products by gel electrophoresis.

15. The method of claim 13 wherein the analyzing of the amplified products comprises detecting the size of the amplified products by capillary electrophoresis.

16. The method of claim 13 wherein the analyzing of the amplified products comprises detecting the size of the amplified products with real time PCR.

17. The method of claim 1 wherein the first and second cultivar are different cultivars of the same cotton species.

18. The method of claim 17 wherein the cotton species is selected from the group consisting of G. barbadense, G. hirsutum, G. arboreum, or G. herbaceum.

19. A method for verifying a textile article comprising:

collecting cotton fibers from a textile item;

extracting genomic DNA from the cotton fibers collected;

amplifying said DNA with PCR-based techniques;

analyzing the amplified product to distinguish between a first cotton cultivar and a second cotton cultivar;

identifying the cotton fibers from the textile item as belonging to the first cotton cultivar or to the second cotton cultivar; and

determining from the identified cotton cultivar if the textile article is authentic.

20. The method of claim 19 wherein first cotton cultivar is from the species G. barbadense.

21. The method of claim 20 wherein the second cotton cultivar is from the species is G. barbadense.

22. The method of claim 19 wherein the amplifying further comprises;

using at least one set of specific primers which targets sequence polymorphism between the first cotton cultivar and the second cotton cultivar.

23. The method of claim 21 wherein amplifying further comprises;

using at least one set of specific primers which targets sequence polymorphism between G. barbadense cultivars.

24. The method of claim 23 wherein the targeted sequence of the first cotton cultivar and the second cotton cultivar have different detectable physical characteristics.

25. The method of claim 24 wherein the detectable physical characteristic of the targeted sequence is a different sequence length for the first cotton cultivar compared the second cotton cultivar.

26. The method of claim 19 wherein the genomic DNA extracted from the cotton fibers comprises chloroplast DNA.

27. The method of claim 23 wherein the detectable physical characteristic of the targeted sequence is a different sequence length.

28. The method of claim 27 wherein the analyzing of the amplified products comprises detecting the size of the amplified products by gel electrophoresis.

29. The method of claim 27 wherein the analyzing of the amplified products comprises detecting the size of the amplified products by capillary electrophoresis.

30. A kit for determining the cotton cultivar utilized in a textile item, comprising:

a sample collection tube for placing mature cotton fibers obtained from the textile item;

a DNA extraction solution;

at least one set of specific primers to distinguish between a first cotton cultivar and a second cotton cultivar; and

a PCR amplification buffer solution.

31. The kit of claim 30 may further comprise a PCR instrument.

32. The kit of claim 30 further comprising an internal control for comparing the size of the amplified product.

33. The kit of claim 30 further comprising a capillary electrophoresis device.